Hai-Bo Wang, Qiu-Hua Mo, Ze Yang*
...leic acid sequence-based amplification (NASBA) and molecular beacon technique. Recently there have been two major progresses in the field, in which RT-NASBA technique was applied for the detection of diarrhoea related viruses. This mini review overviewed the basic principle of RT-NASBA and discussed the significance of these new findings....
Usman Ijaz1, Muhammad Sajid Tahir2, Khalid Abdul Majeed3*, Shahid Iqbal4, Iffat Huma5, S. Firyal6, Ijaz Ahmed7, Shahid Chohan8 and Aamir Riaz Khan9
...Kashmir, Pakistan. After amplification with specific oligos, the amplicons of partial region of COI were subjected to sequencing, and then observed for single nucleotide polymorphisms (SNPs). SNPs were observed on three loci of the COI gene as compared to reference sequence of Panthera pardus pardus. The specimens found in Pakistan were found to be closely related to Panthera pardus orientalis (Amur Leopard in South eastern Russia) and Panther...
Ali Raza Awan*, Sehrish Firyal, Muhammad Tayyab, Lala Rukh, M. Zia ul Haq, Shagufta Saeed and Muhammad Wasim
...solated and utilized for amplification and DNA sequencing of Cytb gene. The Phylogenetic analysis of the Cytb gene indicated that the Pakistani wild Rose ringed Parakeet was mono-phyletically claded with P. k. manillensis with a sequence similarity of 99.37%. Comparative analysis indicated 4 Single Nucleotide Polymorphism (SNPs) sites in the Pakistani wild Rose-ringed parakeet’s Cytb gene which tags the peculiarity to the Pakistani wild Rose-ringed parak...
Shazia Irfan1*, Asima Rani1, Muhammad Arshad2 and Razia Bashir1
...llele specific PCR based amplification strategy. The serum samples were analyzed for determination of metals and minerals through Atomic absorption spectrophotometer (AA 6600 Shimadzu). The statistical analysis indicated that non-significant association existed between SOD1 (rs2070424) gene polymorphism and RA (p>0.05). Results of HWE estimation indicate that allele frequencies were not deviant from HWE in RA group. The results of present study indicates th...
Faiz Muhammad,1, 2,*, Zhang Zhi-Feng,1, Shao Ming-Yu1 and Muhammad Shafi3
...has been cloned by rapid amplification of cDNA (RACE) and anchored PCR method. The full length of the (LvCypA) had 855 bp, containing 495 bp of open reading frame (ORF), encoding 164 amino acids with an estimated molecular mass of 17.6 kDa. The LvCypA has four β- strands. Tissue distribution of the LvCypA after real time analysis revealed that expression is in order of muscle, gill, lymphoid organ and hepatopancrease.
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Nagwan F. Zahran, Ali F. Hamza and Rehab M. Sayed*
...f others in the RAPD-PCR amplification patterns of the irradiated insects.
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Zafar Iqbal1* and Muhammad Khurshid2
...ing various forms of PCR amplification inhibitors. In current study, IC-PCR was used for the detection of Tomato leaf curl New Delhi virus (ToLCNDV) from plant extracts of Nicotiana benthamiana plants inoculated with ToLCNDV. To immunocapture the ToLCNDV, two antisera raised against Tomato yellow leaf curl virus-coat protein (TYLCV-CP) and African cassava mosaic virus-coat protein (ACMV-CP) were used. However, immunocapture of ToLCNDV could only ...
Tausif Ahmad1, Iahtasham Khan2, Saddaf Razzaq1, Saeed-ul-Hassan Khan1,* and Raheela Akhtar3
... genus specific rPCR amplification occurred in all the 5 seropositive samples. In the species-specific rPCR B. abortus was detected in all the samples.
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Matiyar Rahaman Khan1,*, Sabyasachi Pal2, Ghule Tushar Manohar2, Somnath Bhattacharyya3, Amit Singh4, Prahlad Sarkar5 and Samuel Lalliansanga6
...a-esterase phenotype and amplification of rDNA and partial sequence homology of ITS-1and ITS-2. Host differential tests confirmed the race 2 of M. incognita infecting passion fruit. Pathogenic relationship with the passion fruit was proved through inoculation studies and inoculated plants also produced almost similar above-ground symptoms and profuse root galling. The pathogenic potential of M. incognita on passion fruit was determined; the minim...
Nirbhay Kushwaha, Achuit K Singh, Brotati Chattopadhyay and Supriya Chakraborty
Recent advances in geminivirus detection and future perspectives
...etection, rolling circle amplification using the bacteriophage 29 DNApolymerase allows for a reliable diagnosis of geminiviruses and presumably all viruses with small single-stranded circular DNA genomes. The results show the efficiency of this technique in characterizing viral DNA components of several geminiviruses from experimental and natural host plant sources. Nucleotide sequence data offers identification of viral molecule to strain/species level for ac...
Ghulam Abbas1,Asif Nadeem1,*, Masroor Ellahi Babar2, Tanveer Hussain2, Muhammad Sajid Tahir3, Wasim Shehzad1, Rajput Zahid Iqbal3, Muhammad Tayyab1 and Maryam Javed1
...tion and quantification, amplification of gene was done by polymerase chain reaction. PCR products were sequenced bi-directionally, aligned and single nucleotide polymorphisms were identified. Bioinformatics tools were applied for construction of phylogenetic tree and genetic diversity analysis. Lower genetic diversity was observed. The finding of this research is prerequisite for future research.
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Furqan Sabir1, Muhammad Tayyab1,*, Bushra Muneer2, Abu Saeed Hashmi1, Ali Raza Awan1, Naeem Rashid3, Muhammad Wasim1 and Sehrish Firyal1
....The PCR resulted in the amplification of 1.8 kb phytase gene. SDS-PAGE confirmed the size of recombinant protein as 70 kDa. The optimization studies demonstrated the maximal production of recombinant phytase, when the recombinant cells were induced with 1.4 mM IPTG with the post induction time of 6 hours. PHYTN showed maximal activity at 80°C in 50 mM sodium acetate buffer pH 6. Presence of Fe3+ or Cu2+ showed an enhancing...
Tahira Kamal1*, Saeed-Ul-Hassan Khan2, Amir Bin Zahoor3, Khalid Naeem3, Muhammad Naeem Riaz1, Siddra Tayyab Akhtar4, Ghulam Muhammad Ali1*
Ayesha Bibi*, Musharaf Ahmad and Shaukat Hussain
... and CMM-6 were used for amplification of 614bp product from pat-1 gene of bacterial plasmid. The whole bacterial DNA was used as template, extracted with standard procedures using commercially available kit i.e. protinase-K (sigma). 27 out of 34 isolates were confirmed as Cmm having 614bp band. Black nightshade (Solanum nigrum L) was the only weed from which pathogen was consistently isolated. From Northern KP, a total of nine weeds including; Solanum nigrum ...
Lin Wang1,2, Ye Gong3, Kelin Chen1, Haitao Wang3 and Xianguo Lyu1,*
...tested the cross-species amplification of 90 microsatellite loci developed for eight other species. Eleven out of them were successfully amplified and polymorphic with 2-10 alleles. The observed heterozygosities ranged from 0.481 to 0.827 and the polymorphic information content ranged from 0.373 to 0.775. Significant linkage disequilibrium was found only between two markers. These microsatellites could be used to enhance our understanding of genetic informatio...
Rashid Minhas*, Iftikhar Ahmad Khan**, Faisal Saeed Awan**, Lal Hussain Akhtar*, Syed Awais Sajid Shah* and M. Shahjhan Bukhari*
ESTIMATION OF GENETIC DIVERSITY AND HYBRID IDENTIFICATION IN AMERICAN COTTON (GOSSYPIUM HIRSUTUM L.) BY PCR BASED RAPD ANALYSIS
...APD primers used for DNA amplification, 14 primers produced polymorphic bands, while 12 produced monomorphic bands. These 14 primers yielded total 96 RAPD markers, out of which only 40 were found polymorphic which resulted in 41.66% polymorphism among the genotypes studied. The number of RAPD markers produced ranged from 4 to 12. Average number of 6.40 bands per primer was observed. On the whole, 87.50% genetic identity was found in material under investigatio...
Selçuk Kaplan1,* and Sertaç Atalay2
.../i> gene was detected by amplification of the 294 bp region using specific primers and cleavage with the BfoI enzyme. Allele frequencies of A and B were found with 0.915 and 0.085 respectively. The genotype frequencies of IGF1 gene were 0.85 (AA), 0.13 (AB) and 0.02 (BB). The SNP in the exon 3 of the LEP gene was detected by amplification of the 494 bp region using specific primers and cleavage with the ...
Aitezaz Ahsan1*, Muhammad Usman1, Ihsan Ullah2, Aamer Bin Zahur3 and Adnan Rasheed Malik4
...loop mediated isothermal amplification over the last decade. These diagnostic assays vary in their sensitivity, specificity, reliability and reproducibility. Although these assays are detecting PPRV with precision but there is need to develop some cost effective diagnostic assays which would be applicable in the field conditions and in the developing countries like Pakistan.
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Amna Kausar1,2, Sana Anwar1, Naila Siddique2, Safia Ahmed1 and Javid Iqbal Dasti1,*
... specific anti-sera. PCR amplification was done by using N2 specific primers that confirmed 100% of the matrix (M) gene positive isolates as subtype H9N2. This is the first report from Pakistan that confirms prevalence of AIV H9N2 among different bird species across various regions of the country. AIV remains a pandemic threat therefore vigilance for routine AIV surveillance programs and improved vaccination strategies are highly desirable.
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